Linker peptides and polypeptides comprising same
View Patent ↗The invention is based, at least in part, on the finding that linker peptides which lack the amino acid sequence GSG reduce or eliminate the addition of posttranslational modifications to the polypeptides which comprise them. More specifically, the novel linker peptides disclosed herein reduce the ability of enzymes to link carbohydrate adducts to polypeptides comprising these linker peptides, e.g., reduce the ability of xylosyltransferase to link xylose to polypeptides. These novel linker peptides, molecules comprising same, and methods of their use are described.
1. A polypeptide comprising a linker peptide, wherein the linker peptide lacks the sequence GSG and comprises an amino acid sequence selected from the group consisting of:
(a) (GGGGA) 2 GGGGS (SEQ ID NO:8),
(b) (GGGGQ) 2 GGGGS (SEQ ID NO:5),
(c) (GGGPS) 2 GGGGS (SEQ ID NO:6), and
(d) GGGGS(PGGGS) 2 (SEQ ID NO:7).
2. The polypeptide of claim 1 , wherein the linker peptide consists of an amino acid sequence selected from the group consisting of:
(a) (GGGGA) 2 GGGGS (SEQ ID NO:8);
(b) (GGGGQ) 2 GGGGS (SEQ ID NO:5);
(c) (GGGPS) 2 GGGGS (SEQ ID NO:6); and
(d) GGGGS(PGGGS) 2 (SEQ ID NO:7).
3. The polypeptide of claim 1 , wherein the linker peptide is genetically fused to an Fc moiety.
4. The polypeptide of claim 1 , wherein the linker peptide is interposed between two polypeptide domains, wherein at least one of the polypeptide domains comprises a VH domain, a VL domain, a scFv molecule, an Fc moiety, a receptor or extracellular domain thereof, an Fab, and a receptor binding portion of a ligand, an enzyme, a growth factor, an interleukin, a cytokine, or a chemokine.
5. The polypeptide of claim 3 , wherein the Fc moiety is an Fc region.
6. The polypeptide of claim 3 , wherein the Fc moiety is an scFc region.
7. The polypeptide of claim 1 , wherein the polypeptide is a bispecific antibody molecule.
8. A composition comprising the polypeptide of claim 1 and a pharmaceutically acceptable carrier.
9. A binding molecule comprising an scFv moiety and an Fc moiety, wherein the scFv moiety and the Fc moiety are genetically linked by a linker peptide, wherein the linker peptide comprises an amino acid sequence selected from the group consisting of:
(a) (GGGGA) 2 GGGGS (SEQ ID NO: 8),
(b) (GGGGQ) 2 GGGGS (SEQ ID NO:5),
(c) (GGGPS) 2 GGGGS (SEQ ID NO:6), and
(d) GGGGS(PGGGS) 2 (SEQ ID NO:7),
and wherein the linker peptide lacks the sequence GSG.
10. The binding molecule of claim 9 , wherein the linker peptide consists of an amino acid sequence selected from the group consisting of:
(a) (GGGGA) 2 GGGGS (SEQ ID NO:8),
(b) (GGGGQ) 2 GGGGS (SEQ ID NO:5),
(c) (GGGPS) 2 GGGGS (SEQ ID NO:6), and
(d) GGGGS(PGGGS) 2 (SEQ ID NO:7).
11. A composition comprising the binding molecule of claim 9 and a pharmaceutically acceptable carrier.
12. An scFv molecule comprising a VH and a VL region, wherein the VH and VL region are genetically linked by a linker peptide, wherein the linker peptide comprises an amino acid sequence selected from the group consisting of:
(a) (GGGGA) 2 GGGGS (SEQ ID NO:8);
(b) (GGGGQ) 2 GGGGS (SEQ ID NO:5);
(c) (GGGPS) 2 GGGGS (SEQ ID NO:6);
(d) GGGGS(PGGGS) 2 (SEQ ID NO:7);
(e) (GGGGA) 3 (SEQ ID NO:12); and
(f) (GGGGA) 4 (SEQ ID NO:13),
and wherein the linker peptide lacks the sequence GSG.
13. The scFv molecule of claim 12 , wherein the linker peptide consists of an amino acid sequence selected from the group consisting of:
(a) (GGGGA) 2 GGGGS (SEQ ID NO:8);
(b) (GGGGQ) 2 GGGGS (SEQ ID NO:5);
(c) (GGGPS) 2 GGGGS (SEQ ID NO:6);
(d) GGGGS(PGGGS) 2 (SEQ ID NO:7);
(e) (GGGGA) 3 (SEQ ID NO:12); and
(f) (GGGGA) 4 (SEQ ID NO:13).
14. A composition comprising the scFv molecule of claim 12 and a pharmaceutically acceptable carrier.
15. A binding molecule comprising an scFv moiety which comprises a VH and a VL region and an Fc moiety, wherein the scFv moiety and the Fc moiety are genetically linked by a first linker peptide consisting of an amino acid sequence selected from the group consisting of:
(a) (GGGGA) 2 GGGGS (SEQ ID NO: 8);
(b) (GGGGQ) 2 GGGGS (SEQ ID NO:5),
(c) (GGGPS) 2 GGGGS (SEQ ID NO:6), and
(d) GGGGS(PGGGS) 2 (SEQ ID NO:7),
and the VH and the VL region are genetically linked by a second linker peptide consisting of an amino acid sequence selected from the group consisting of:
(e) (GGGGA) 3 (SEQ ID NO:12); and
(f) (GGGGA) 4 (SEQ ID NO:13).
16. A composition comprising the binding molecule of claim 15 and a pharmaceutically acceptable carrier.
17. A linker peptide selected from the group consisting of:
(a) a linker peptide comprising the amino acid sequence (GGGGA) 2 GGGGS (SEQ ID NO:8);
(b) a linker peptide comprising the amino acid sequence (GGGGQ) 2 GGGGS (SEQ ID NO:5);
(c) a linker peptide comprising the amino acid sequence (GGGPS) 2 GGGGS (SEQ ID NO:6);
(d) a linker peptide comprising the amino acid sequence GGGGS(PGGGS) 2 (SEQ ID NO:7);
(e) a linker peptide consisting of the amino acid sequence (GGGGA) 2 GGGGS (SEQ ID NO:8);
(f) a linker peptide consisting of the amino acid sequence (GGGGQ) 2 GGGGS (SEQ ID NO:5);
(g) a linker peptide consisting of the amino acid sequence (GGGPS) 2 GGGGS (SEQ ID NO:6); and
(h) a linker peptide consisting of the amino acid sequence GGGGS(PGGGS) 2 (SEQ ID NO:7).
18. The binding molecule of claim 9 , wherein the Fc moiety is an Fc region.
19. The binding molecule of claim 9 , wherein the Fc moiety is an scFc region.
20. The binding molecule of claim 9 , wherein the binding molecule is a bispecific molecule.
21. The binding molecule of claim 9 , wherein the scFv moiety comprises a VH and a VL region, wherein the VH and VL region are genetically linked by a linker peptide comprising the amino acid sequence (GGGGA) 4 (SEQ ID NO:13).
22. The polypeptide of claim 2 , wherein the linker peptide is genetically fused to an Fc moiety.
23. The polypeptide of claim 2 , wherein the linker peptide is interposed between two polypeptide domains, wherein at least one of the polypeptide domains comprises a VH domain, a VL domain, a scFv molecule, an Fc moiety, a receptor or extracellular domain thereof, an Fab, and a receptor binding portion of a ligand, an enzyme, a growth factor, an interleukin, a cytokine, or a chemokine.
24. The polypeptide of claim 22 , wherein the Fc moiety is an Fc region.
25. The polypeptide of claim 22 , wherein the Fc moiety is an scFc region.
26. The polypeptide of claim 2 , wherein the polypeptide is a bispecific antibody molecule.
27. A composition comprising the polypeptide of claim 2 and a pharmaceutically acceptable carrier.